BaZiBuShen inhibition of oocyte ferroptosis in primordial follicles through activation of NF2-YAP pathway for the treatment of chemotherapy-induced premature ovarian insufficiency.

ETHNOPATHOLOGICAL RELEVANCE

BaZiBuShen (BZBS) is an innovative, patented traditional Chinese medicine known for its kidney-tonifying and anti-aging effects. It contains active ingredients such as flavonoids and amino acid analogues, which have anti-inflammatory and antioxidant properties, and is used to alleviate symptoms of "kidney essence" deficiency.

AIM OF THE STUDY

This study evaluated the efficacy of BZBS on cyclophosphamide (CTX)-induced premature ovarian insufficiency (POI) and explored its possible mechanism of action.

METHODS

POI models in rats were established using CTX to assess the therapeutic effects of BZBS. HPLC-MS was used to analyze the components, while ELISA was adopted to determine the serum hormone levels. Ovarian morphology and number of follicles were evaluated by H&E staining. The ultrastructure of mitochondria was examined by TEM. The expression levels of proteins related to ferroptosis and the NF2-YAP signalling pathway were analysed using immunohistochemistry, immunofluorescence, and Western blotting.

RESULTS

In CTX-induced POI rats, BZBS treatment effectively restored ovarian weight, while simultaneously decreasing serum FSH and LH levels and increasing E levels. Histological analysis of the ovaries revealed that BZBS significantly increased the number of primordial, growing, and mature follicles, as well as reducing the number of atretic follicles. Furthermore, BZBS treatment mitigated ferroptosis by decreasing key markers Fe, TFR, ACSL4, and restoring the levels of GSH and GPX4. Additionally, BZBS modulated the expression of critical proteins involved in ferroptosis and cell signalling pathways. Specifically, it down-regulated p-RB1, while up-regulating SLC7A11 and RB1. Moreover, BZBS upregulates NF2 while downregulating YAP expression and its nuclear translocation, thereby regulating the NF2-YAP signaling pathway involved in ferroptosis.

CONCLUSIONS

In a CTX-induced POI rat model, BZBS effectively restores hormonal levels, mitigates ovarian damage, and curbs excessive primordial follicle activation. It also modulates ferroptosis-related protein expression, activates the NF2-YAP pathway, and could provide a potential therapeutic approach for POI.